This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Fuller, M. E.
Right arrow Articles by DeFlaun, M. F.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Fuller, M. E.
Right arrow Articles by DeFlaun, M. F.
Agricola
Right arrow Articles by Fuller, M. E.
Right arrow Articles by DeFlaun, M. F.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, October 2000, p. 4486-4496, Vol. 66, No. 10
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Development of a Vital Fluorescent Staining Method for Monitoring Bacterial Transport in Subsurface Environments

Mark E. Fuller,1,* Sheryl H. Streger,1 Randi K. Rothmel,1 Brian J. Mailloux,2 James A. Hall,2 Tullis C. Onstott,2 James K. Fredrickson,3 David L. Balkwill,4 and Mary F. DeFlaun1

Envirogen, Inc., Princeton Research Center, Lawrenceville, New Jersey 086481; Department of Geosciences, Princeton University, Princeton, New Jersey 085442; Pacific Northwest National Laboratory, Richland, Washington 993383; and Department of Biological Science, Florida State University, Tallahassee, Florida 323064

Received 3 May 2000/Accepted 23 July 2000

Previous bacterial transport studies have utilized fluorophores which have been shown to adversely affect the physiology of stained cells. This research was undertaken to identify alternative fluorescent stains that do not adversely affect the transport or viability of bacteria. Initial work was performed with a groundwater isolate, Comamonas sp. strain DA001. Potential compounds were first screened to determine staining efficiencies and adverse side effects. 5-(And 6-)-carboxyfluorescein diacetate, succinimidyl ester (CFDA/SE) efficiently stained DA001 without causing undesirable effects on cell adhesion or viability. Members of many other gram-negative and gram-positive bacterial genera were also effectively stained with CFDA/SE. More than 95% of CFDA/SE-stained Comamonas sp. strain DA001 cells incubated in artificial groundwater (under no-growth conditions) remained fluorescent for at least 28 days as determined by epifluorescent microscopy and flow cytometry. No differences in the survival and culturability of CFDA/SE-stained and unstained DA001 cells in groundwater or saturated sediment microcosms were detected. The bright, yellow-green cells were readily distinguished from autofluorescing sediment particles by epifluorescence microscopy. A high throughput method using microplate spectrofluorometry was developed, which had a detection limit of mid-105 CFDA-stained cells/ml; the detection limit for flow cytometry was on the order of 1,000 cells/ml. The results of laboratory-scale bacterial transport experiments performed with intact sediment cores and nondividing DA001 cells revealed good agreement between the aqueous cell concentrations determined by the microplate assay and those determined by other enumeration methods. This research indicates that CFDA/SE is very efficient for labeling cells for bacterial transport experiments and that it may be useful for other microbial ecology research as well.

* Corresponding author. Mailing address: Envirogen, Inc., Princeton Research Center, 4100 Quakerbridge Road, Lawrenceville, NJ 08648. Phone: (609) 936-1815, ext. 169. Fax: (609) 936-9221. E-mail: fuller{at}envirogen.com.

Applied and Environmental Microbiology, October 2000, p. 4486-4496, Vol. 66, No. 10
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Busscher, H. J., van der Mei, H. C. (2006). Microbial Adhesion in Flow Displacement Systems. Clin. Microbiol. Rev. 19: 127-141 [Abstract] [Full Text]  
  • Fuller, M. E., Mailloux, B. J., Streger, S. H., Hall, J. A., Zhang, P., Kovacik, W. P., Vainberg, S., Johnson, W. P., Onstott, T. C., DeFlaun, M. F. (2004). Application of a Vital Fluorescent Staining Method for Simultaneous, Near-Real-Time Concentration Monitoring of Two Bacterial Strains in an Atlantic Coastal Plain Aquifer in Oyster, Virginia. Appl. Environ. Microbiol. 70: 1680-1687 [Abstract] [Full Text]  
  • Mailloux, B. J., Fuller, M. E. (2003). Determination of In Situ Bacterial Growth Rates in Aquifers and Aquifer Sediments. Appl. Environ. Microbiol. 69: 3798-3808 [Abstract] [Full Text]  
  • Dong, H., Rothmel, R., Onstott, T. C., Fuller, M. E., DeFlaun, M. F., Streger, S. H., Dunlap, R., Fletcher, M. (2002). Simultaneous Transport of Two Bacterial Strains in Intact Cores from Oyster, Virginia: Biological Effects and Numerical Modeling. Appl. Environ. Microbiol. 68: 2120-2132 [Abstract] [Full Text]  
  • Harvey, R. W., Mayberry, N., Kinner, N. E., Metge, D. W., Novarino, F. (2002). Effect of Growth Conditions and Staining Procedure upon the Subsurface Transport and Attachment Behaviors of a Groundwater Protist. Appl. Environ. Microbiol. 68: 1872-1881 [Abstract] [Full Text]  
  • Johnson, W. P., Zhang, P., Gardner, P. M., Fuller, M. E., DeFlaun, M. F. (2001). Evidence for Detachment of Indigenous Bacteria from Aquifer Sediment in Response to Arrival of Injected Bacteria. Appl. Environ. Microbiol. 67: 4908-4913 [Abstract] [Full Text]  
  • Mogull, S. A., Runyen-Janecky, L. J., Hong, M., Payne, S. M. (2001). dksA Is Required for Intercellular Spread of Shigella flexneri via an RpoS-Independent Mechanism. Infect. Immun. 69: 5742-5751 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»